Trifludimoxazin is a new herbicide that inhibits protoporphyrinogen oxidase and is targeted for commercial market introduction in North America, South America, and Asia. It will be available both as a standalone product and in a 1:2 mixture with saflufenacil. The herbicide is intended for use in preplant burndown (PPBD) and preemergence applications in cereal, corn (Zea mays L.), soybean [Glycine max (L.) Merr.], and pulse crops to control a variety of annual broadleaf and grass weed species. Additionally, it is planned to be used in tree crops, oil-palm (Elaeis quineensis Jacq.), and non-crop areas.

In this study, we meticulously evaluated the performance and effectiveness of both the standalone herbicide and the innovative mixture concept in combating prevalent weeds commonly encountered in corn and soybean fields. Our findings revealed that both products exhibited exceptional efficacy, significantly reducing the presence of these troublesome weeds. Furthermore, the mixture concept not only demonstrated commendable soil mobility but also showcased impressive residual activity, positioning it as a powerful tool for sustainable weed control. These promising effects are further substantiated by our comprehensive ADME (Adsorption-Distribution-Metabolism-Extraction) studies, which provide insight into the behavior and longevity of the herbicides in the agricultural ecosystem.

Protoporphyrinogen oxidase (PPO)-inhibiting herbicides remain an important and useful chemistry 60 yr after their first introduction. In this review, based on topics introduced at the Weed Science Society of America 2021 symposium titled “A History, Overview, and Plan of Action on PPO Inhibiting Herbicides,” we discuss the current state of PPO-inhibiting herbicides. Renewed interest in the PPO-inhibiting herbicides in recent years, due to increased use and increased cases of resistance, has led to refinements in knowledge regarding the mechanism of action of PPO inhibitors. Herein we discuss the importance of the two isoforms of PPO in plants, compile a current knowledge of target-site resistance mechanisms, examine non–target site resistance cases, and review crop selectivity mechanisms. Consistent and reproducible greenhouse screening and target-site mutation assays are necessary to effectively study and compare PPO-inhibitor resistance cases. To this end, we cover best practices in screening to accurately identify resistance ratios and properly interpret common screens for point mutations. The future of effective and sustainable PPO-inhibitor use relies on development of new chemistries that maintain activity on resistant biotypes and the promotion of responsible stewardship of PPO inhibitors both new and old. We present the biorational design of the new PPO inhibitor trifludimoxazin to highlight the future of PPO-inhibitor development and discuss the elements of sustainable weed control programs using PPO inhibitors, as well as how responsible stewardship can be incentivized. The sustained use of PPO inhibitors in future agriculture relies on the effective and timely communication from mode of action and resistance research to agronomists, Extension workers, and farmers.

Three dose-response trials were performed in 2020 and 2021 to determine the tolerance of two Jack O’Lantern pumpkin cultivars to fomesafen applied preemergence at two Indiana locations: the Southwest Purdue Agricultural Center (SWPAC) and the Pinney Purdue Agricultural Center (PPAC). The experiment was a split-plot arrangement in which the main plot was the fomesafen rate of application (0, 280, 560, 840, and 1,220 g ai ha–1), and the subplot was the pumpkin cultivar (‘Bayhorse Gold’ and ‘Carbonado Gold’). As the fomesafen rate increased from 280 to 1,120 g ha–1, the predicted pumpkin emergence decreased from 85% to 25% of the nontreated control at SWPAC-2020, but only from 99% to 74% at both locations in 2021. The severe impact on emergence at SWPAC-2020 was attributed to rainfall. Visible injury included bleaching and chlorosis due to the herbicide splashing from the soil surface onto the leaves and included stunting, but injury was transient. As the fomesafen rate increased from 280 to 1,120 g ha–1, the predicted marketable orange pumpkin yield decreased from 95% to 24% of the nontreated control at SWPAC-2020 and 98% to 74% at PPAC-2021. Similarly, the predicted marketable orange pumpkin fruit number decreased from 94% to 21% at SWPAC-2020 and 98% to 74% at PPAC-2021. Fomesafen rate did not affect marketable orange pumpkin yield and fruit number at SWPAC-2021 and marketable orange pumpkin fruit weight at any location-year. Overall, the fomesafen rate of 280 g ha–1 was safe for use preemergence in the pumpkin cultivars ‘Bayhorse Gold’ and ‘Carbonado Gold’ within one day after planting, but there is a risk of increased crop injury with increasing rainfall.

False-green kyllinga is a problematic C4 perennial Cyperaceae species. Previous research examined herbicide efficacy but has not integrated nonchemical practices with herbicides. Replicate field experiments were conducted to evaluate late-summer postemergence applications of herbicides in combination with turf-type tall fescue interseeding for false-green kyllinga control. Seven herbicide treatments and a nontreated control formed a complete factorial, with tall fescue interseeding or no seeding (eight-by-two factorial). Six herbicide treatments consisted of single postemergence applications of halosulfuron-methyl (70 g ha–1), imazosulfuron (420 g ha–1), and sulfentrazone + carfentrazone (280 + 30 g ha–1) applied 4 wk before tall fescue interseeding (WBS) and the day of interseeding. Glyphosate (220 g ae ha–1) applied the day of interseeding was the seventh herbicide treatment. Tall fescue was interseeded in September, and false-green kyllinga control was evaluated the following summer. In combination with tall fescue interseeding, imazosulfuron and halosulfuron applied 4 WBS as well as glyphosate applied the day of interseeding controlled false-green kyllinga better than all other treatments. In July, imazosulfuron and halosulfuron treatments applied 4 WBS and combined with interseeding had 1% to 6% false-green kyllinga cover, respectively, compared to 28% and 62% cover, respectively, without interseeding. Interseeding alone controlled false-green kyllinga <50%. Imazosulfuron, halosulfuron, and sulfentrazone + carfentrazone applied the day of seeding severely injured emerging tall fescue seedlings and reduced turfgrass quality the following spring. Applying imazosulfuron or halosulfuron in late summer and interseeding turf-type tall fescue 4 wk later or applying glyphosate the day of seeding are effective strategies for postemergence false-green kyllinga control. Integrating herbicides and the cultural practice of tall fescue interseeding provided more false-green kyllinga control than either practice alone.

Trifludimoxazin, a new protoporphyrinogen oxidase–inhibiting herbicide, is being evaluated for possible use as a soil-residual active herbicide treatment in cotton for control of small-seeded annual broadleaf weeds. Laboratory and greenhouse studies were conducted to compare vertical mobility and cotton tolerance of trifludimoxazin to flumioxazin and saflufenacil, which are two currently registered protoporphyrinogen oxidase–inhibiting herbicides for use in cotton, in three West Texas soils. Vertical soil mobility of trifludimoxazin was similar to flumioxazin in Acuff loam and Olton loam soils, but was more mobile than flumioxazin in the Amarillo loamy sand soil. The depth of trifludimoxazin movement after a 2.5-cm irrigation event ranged from 2.5 to 5.0 cm in all soils, which would not allow for crop selectivity based on herbicide placement, because ideal cotton seeding depth is from 0.6 to 2.54 cm deep. Greenhouse studies indicated that PRE treatments were more injurious than the 14 d preplant treatment when summarized across soils for the three herbicides (43% and 14% injury, respectively). No differences in visual cotton response or dry weight was observed after trifludimoxazin preplant as compared with the nontreated control within each of the three West Texas soils and was similar to the flumioxazin preplant across soils. On the basis of these results, a use pattern for trifludimoxazin in cotton may be established with the use of a more than 14-d preplant restriction before cotton planting.

We conducted research to evaluate various herbicides for POST false-green kyllinga control in cool-season turfgrass (primarily creeping bentgrass). In a preliminary evaluation, single and sequential applications of halosulfuron-methyl (70 g ai ha−1), mesotrione (175 g ai ha−1), and sulfentrazone (140 g ai ha−1), as well as a single application of imazosulfuron (740 g ai ha−1), were evaluated in New Jersey. Imazosulfuron and sequential applications of halosulfuron-methyl controlled false-green kyllinga >93% at 9 and 18 wk after initial treatment (WAIT). Sulfentrazone and mesotrione controlled false-green kyllinga <50%. Additional experiments were conducted to evaluate single and sequential applications of halosulfuron-methyl (70 g ha−1), imazosulfuron (420 and 740 g ha−1), and sulfentrazone (140 g ha−1) in New Jersey and Indiana at two locations in each state. At 12 WAIT, imazosulfuron generally controlled false-green kyllinga more effectively than other treatments at all locations. Sequential applications of imazosulfuron controlled false-green kyllinga 100% at 12 WAIT. Halosulfuron-methyl was less effective in Indiana than in New Jersey. Sulfentrazone controlled false-green kyllinga <40% at 12 WAIT. This research demonstrates that imazosulfuron is more effective than halosulfuron-methyl and sulfentrazone for POST false-green kyllinga control in cool-season turf.

Palmer amaranth (Amaranthus palmeri S. Watson), a dioecious summer annual species, is one of the most troublesome weeds in U.S. cropping systems. The evolution of resistance to protoporphyrinogen oxidase inhibitors in A. palmeri biotypes is a major cause of concern to soybean [Glycine max (L.) Merr.] and cotton (Gossypium hirsutum L.) growers in the midsouthern United States. The objective of this study was to confirm and characterize the non–target site mechanism in a fomesafen-resistant accession from Randolph County, AR (RCA). A dose–response assay was conducted to assess the level of fomesafen resistance, and based on the GR50 values, the RCA accession was 18-fold more resistant to fomesafen than a susceptible (S) biotype. A TaqMan allelic discrimination assay and sequencing of the target-site genes PPX2 and PPX1 revealed no known or novel target-site mutations. An SYBR Green assay indicated no difference in PPX2 gene expression between the RCA and S biotypes. To test whether fomesafen resistance is metabolic in nature, the RCA and the S biotypes were treated with different cytochrome P450 (amitrole, piperonyl butoxide [PBO], malathion) and glutathione S-transferase (GST) (4-chloro-7-nitrobenzofurazan [NBD-Cl]) inhibitors, either alone or in combination with fomesafen. Malathion followed by (fb) fomesafen in RCA showed the greatest reduction in survival (67%) and biomass (86%) compared with fomesafen alone (45% and 66%, respectively) at 2 wk after treatment. Interestingly, NBD-Cl fb fomesafen also resulted in low survival (35%) compared with the fomesafen-only treatment (55%). Applications of malathion or NBD-Cl preceding fomesafen treatment resulted in reversal of fomesafen resistance, indicating the existence of cytochrome P450– and GST-based non–target site mechanisms in the RCA accession. This study confirms the first case of non–target site resistance to fomesafen in A. palmeri.

Greenhouse-grown soybean cultivars varied in their tolerance to preemergence application of sulfentrazone. The cultivars Ransom, Hutcheson, Kato, Gasoy 17, and Cobb exhibited relatively low tolerance to 0.5 kg ai ha−1 sulfentrazone with 38, 41, 46, 50, and 58% height reduction compared to respective controls. The growth of tolerant cultivars Centennial, Edison, and Hartz 5164 was not affected by this treatment. However, the growth of all cultivars was reduced at the excessive rate of 2.0 kg ha−1 preemergence application of sulfentrazone. No differences in root uptake or translocation of [14C] sulfentrazone were observed between the relatively tolerant and less tolerant cultivars tested. Centennial and Hutcheson cultivars rapidly metabolized sulfentrazone via oxidative degradation of the 3-methyl group on the triazolinone ring of the herbicide. Only 4.7 and 4.9% of the active ingredient remained in the foliage of Hutcheson and Centennial 24 h after treatment, respectively. While there were no differences in Protox inhibition or Proto IX accumulation between the two cultivars, Hutcheson was more sensitive than Centennial to peroxidative stresses induced by either Proto IX accumulation or rose bengal. Therefore, tolerance to sulfentrazone is due to rapid metabolism of the herbicide; however, the intraspecific difference in response to sulfentrazone appears to be due to intrinsic differential tolerance to the herbicide-induced peroxidative stress.

Field experiments were conducted in Platte County, Missouri, during 2006 and 2007 to evaluate PRE, POST, and PRE followed by (fb) POST herbicide programs for the control of glyphosate-resistant waterhemp in soybean. All PRE fb POST treatments resulted in at least 66 and 70% control of glyphosate-resistant waterhemp in 2006 and 2007, respectively. Control of glyphosate-resistant waterhemp was less than 23% with lactofen and acifluorfen in 2006, but at least 64% in 2007. Variability in control likely resulted from differences in trial locations and a population of protoporphyrinogen oxidase (PPO)–resistant waterhemp at the Platte County site in 2006 compared with 2007. In both years, glyphosate resulted in less than 23% control of glyphosate-resistant waterhemp and provided the least control of all herbicide programs. Programs containing PRE herbicides resulted in waterhemp densities of less than 5 plants/m2, whereas the POST glyphosate treatment resulted in 38 to 70 plants/m2. Waterhemp seed production was reduced at least 78% in all PRE fb POST programs, from 55 to 71% in POST programs containing lactofen and acifluorfen and by only 21% in the POST glyphosate treatment. Soybean yields corresponded to the level of waterhemp control achieved in both years, with the lowest yields resulting from programs that provided poorest waterhemp control. PRE applications of S-metolachlor plus metribuzin provided one of the highest net incomes in both years and resulted in $271 to $340/ha greater net income than the glyphosate-only treatment. Collectively, the results from these experiments illustrate the effectiveness of PRE herbicides for the control of glyphosate-resistant waterhemp in glyphosate-resistant soybean and the inconsistency of PPO-inhibiting herbicides or PPO-inhibiting herbicide combinations for the control of waterhemp populations with multiple resistance to glyphosate and PPO-inhibiting herbicides.

Herbicides inhibit biochemical and physiological processes or both with lethal consequences. The target sites of these small molecules are usually enzymes involved in primary metabolic pathways or proteins carrying out essential physiological functions. Herbicides tend to be highly specific for their respective target sites and have served as tools to study these physiological and biochemical processes in plants (Dayan et al. 2010b).

Carfentrazone-ethyl absorption, translocation, and metabolism was determined in Glycine max, Zea mays, and Abutilon theophrasti. Glycine max absorbed greater than 90% of applied carfentrazone-ethyl within 2 h after treatment (HAT) when nonionic surfactant (NIS) or crop oil concentrate (COC) were included in the treatment solution. The addition of 28% urea ammonium nitrate (UAN) did not improve carfentrazone-ethyl absorption in G. max, but in Z. mays and A. theophrasti, UAN combined with NIS or COC increased the rate of carfentrazone-ethyl absorption. Carfentrazone absorption in A. theophrasti 2 HAT was 70% when UAN was combined with NIS or COC compared to 40% with NIS or COC alone; however, 24 HAT, absorption with NIS and COC were similar to treatments with UAN. Carfentrazone-ethyl did not translocate from the treated leaf to other plant parts in Z. mays and only small amounts of radiolabeled product were detected in the rest of the shoots of A. theophrasti (5%) and G. max (12%). Herbicide metabolism in Z. mays and G. max was greater than in A. theophrasti. All three species converted carfentrazone-ethyl to its phytotoxic metabolite carfentrazone-chloropropionic acid; therefore, the parent molecule was considered to be the sum of the ethyl ester and its hydrolysis product. Estimated half-lives of carfentrazone in Z. mays, G. max and A. theophrasti were 1, 7, and 40 h, respectively. The rate of carfentrazone metabolism corresponded to plant sensitivity (sensitivity to carfentrazone: Z. mays<G. max<<A. theophrasti); however, rapid absorption and translocation of carfentrazone may reduce the tolerance of G. max.

Common waterhemp seeds were collected from two Missouri soybean fields where biotypes were not controlled by acifluorfen. Plants grown from these seeds were tested for resistance to the diphenyl ether herbicides acifluorfen and lactofen. Resistance to susceptibility (R/S) ratios, calculated as the ratio of the dose required to inhibit dry weight accumulation by 50% (GR50) in resistant plants to that for susceptible plants, were 9.5 and 11 for the Meadville biotype and 28 and 44 for the Bethel biotype exposed to acifluorfen and lactofen, respectively. Electrolyte leakage assays determined that light-induced lipid peroxidation by acifluorfen was greatest on a control population (Bradford), intermediate for the Meadville biotype, and lowest for the Bethel biotype. Levels of the photodynamic pigment protoporphyrin IX (Proto) accumulating in leaf disks exposed to acifluorfen were much lower in the resistant biotypes than in the susceptible wild type, and the level of Proto accumulation was significantly correlated to the degree of membrane disruption. Although the binding of acifluorfen to protoporphyrinogen oxidase in chloroplasts may have been altered in the resistant biotypes, the molecular and biochemical factors involved in the mechanism of resistance remain to be fully characterized. However, this study establishes that the physiological basis for the evolved resistance to diphenyl ethers in common waterhemp rests on the reduction of Proto accumulation.

A population of waterhemp was identified in Adams County, Illinois, that survived treatment of several acetolactate synthase (ALS) inhibitors and a postemergence (POST) application of lactofen, a protoporphyrinogen oxidase (PPO)–inhibiting herbicide. Greenhouse studies were conducted to quantify the responses of this waterhemp population, designated ACR, to multiple PPO inhibitors and various other herbicides with different sites of action. Resistance ratios were obtained by comparing herbicide dose–response curves between the ACR population and a herbicide-susceptible waterhemp population. The ACR population was resistant to lactofen (23-fold) and to five other PPO-inhibiting herbicides (ranging from 2.2- to 6.2-fold). Furthermore, the ACR waterhemp population was 17,000-fold and 18,000-fold resistant to imazamox and thifensulfuron, respectively, two ALS-inhibiting herbicides. In response to atrazine, a Photosystem II inhibitor, the ACR population was 38-fold resistant. Plants within the ACR waterhemp population survived treatment of a herbicide mixture containing lactofen at 175 g ai ha−1, imazamox at 44 g ae ha−1, and atrazine at 1,000 g ai ha−1. Thus, individual plants—not just the population as a whole—displayed multiple herbicide resistance. The ACR population was not resistant to glyphosate or paraquat. This is the first reported weed population from the United States with resistances to herbicides inhibiting three unique sites of action. Furthermore, this research identifies a significant reduction in the number of POST herbicide options available for waterhemp control in soybean production.

Potato exhibits adequate tolerance to preemergence applications of sulfentrazone at rates up to 0.28 kg ai ha−1. Sulfentrazone also controls several troublesome weeds such as common lambsquarters but may be less effective against jimsonweed. Laboratory experiments were conducted to investigate differential tolerance to root-absorbed [14C]sulfentrazone by potato, common lambsquarters, and jimsonweed. Common lambsquarters and jimsonweed absorption of [14C]sulfentrazone g−1 fresh weight was more than twofold that in potato after 24-h exposure. After 48-h exposure, sulfentrazone absorption by common lambsquarters was nearly twofold that in jimsonweed and nearly threefold that in potato. Sulfentrazone movement from roots to shoots was also greater in common lambsquarters than in jimsonweed and potato after 6 h. Both weed species exhibited nearly a twofold increase in sulfentrazone translocation from roots to shoots compared with potato after 12, 24, and 48 h. Minor differences in sulfentrazone metabolism in roots were noted among species after 6 h. Metabolism in roots and shoots was similar in all species after 12, 24, and 48 h. Because the enzyme on which sulfentrazone acts, protoporphyrinogen oxidase, is located in shoot tissue, translocation to shoots is essential for sulfentrazone toxicity. Therefore, differential root absorption and differential translocation of sulfentrazone from roots to shoots are the proposed primary mechanisms of differential sulfentrazone tolerance among potato, common lambsquarters, and jimsonweed.

Resistance to protoporphyrinogen oxidase (PPO)-inhibiting herbicides in waterhemp has been shown previously to be the result of a unique mechanism. Specifically, a three–base-pair (3-bp) deletion in the PPX2L gene, a gene encoding both plastid- and mitochondria-targeted PPO enzymes, confers herbicide resistance in this species. Furthermore, when this unique mechanism was initially characterized it was presumed that waterhemp contained three PPX genes, PPX1, PPX2S, and PPX2L, and that the resistant biotypes were missing PPX2S. Here, allele testing and examination of genetic sequence data demonstrate that there are likely only two PPX genes in waterhemp, PPX1 and PPX2L. Next, to determine the prevalence of this mechanism of resistance in Illinois waterhemp, we developed an allele-specific polymerase chain reaction (PCR) marker that amplifies only the 3-bp deletion allele, ΔG210, of PPX2L. By utilizing this marker, we show that the ΔG210 PPX2L allele correlated with whole-plant resistance to PPO inhibitors in each of four other waterhemp populations evaluated from Illinois.

A population of common ragweed from Delaware was not controlled in the field by herbicides that inhibit acetolactate synthase (ALS) or protoporphyrinogen oxidase (PPO). Research was conducted to ascertain if this population was resistant to these herbicidal modes of action and, if so, to determine the resistance mechanism(s). Resistance was confirmed by dose-response studies on greenhouse-grown plants with multiple ALS- and PPO-inhibiting herbicides. DNA sequence data revealed that resistance to ALS-inhibiting herbicides was due to the previously reported W574L ALS mutation. To assist in determining the mechanism of resistance to PPO-inhibiting herbicides, an F2 population was derived from a cross between the resistant biotype (Del-R) and a sensitive biotype (DV1-S). This population segregated in the ratio of three resistant : one sensitive when treated with fomesafen, indicating that resistance to PPO-inhibiting herbicides was conferred by a single, (incompletely) dominant, nuclear gene. Sequences of the target-site genes, PPX1 and PPX2, for PPO-inhibiting herbicides were obtained through the screening of a common ragweed cDNA library and subsequent cDNA extension (5′-RACE). Molecular marker analysis with the F2 population revealed that the PPX2 gene cosegregated with resistance to PPO-inhibiting herbicides. A mutation substituting an arginine codon for a leucine codon at a conserved location (R98L) of the PPX2 gene was suspected of being responsible for resistance. By using a transgenic Escherichia coli system, it was demonstrated that the R98L mutation was sufficient to confer resistance to PPO-inhibiting herbicides. The level of resistance to acifluorfen conferred by the R98L mutation in the E. coli system was about 31-fold, similar to the level of resistance seen in the whole-plant dose-response study. Last, a DNA-based assay was developed to identify the presence or absence of the common ragweed PPX2 R98L mutation. The R98L PPX2 mutation is the second mechanism identified for evolved resistance to PPO-inhibiting herbicides.